scholarly journals Diagnosis of invasive fungal infections by a real-time panfungal PCR assay in immunocompromised pediatric patients

Leukemia ◽  
2010 ◽  
Vol 24 (12) ◽  
pp. 2032-2038 ◽  
Author(s):  
C Landlinger ◽  
S Preuner ◽  
L Bašková ◽  
M van Grotel ◽  
N G Hartwig ◽  
...  
Keyword(s):  
Real Time ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Pcr Assay ◽  
Panfungal Pcr

Diagnosis of Invasive Fungal Infections by a Real-Time Panfungal PCR Assay in Pediatric Patients Undergoing Intensive Chemotherapy or Allogeneic Stem Cell Transplantion.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 343-343
Author(s):  
Christine Landlinger ◽  
Lenka Baskova ◽  
Sandra Preuner ◽  
Martine Van Grotel ◽  
Nico G. Hartwig ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Predictive Value ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Immunocompromised Patients ◽  
Pcr Assay ◽  
Highly Sensitive ◽  
Panfungal Pcr

Abstract Abstract 343 Invasive fungal infections are life threatening events in severely immunocompromised patients, and there is urgent need for reliable screening methods facilitating rapid and broad detection of pathogenic fungi. We have established a two-reaction real-time PCR assay permitting highly sensitive detection and quantitative monitoring of more than 80 fungal pathogens, covering a large spectrum of moulds, yeasts and Zygomycetes (European patent No. 06817468.9). To assess the clinical potential of the panfungal real-time PCR assay, more than 600 consecutive specimens from 126 pediatric patients carrying a high risk of invasive fungal infections were analyzed. The results revealed an excellent correlation between PCR positivity and the presence of proven, probable or possible fungal infection according to the criteria of the European Organization for Research and Treatment of Cancer (EORTC), indicating a sensitivity of the assay of 96% (95%CI: 81-99.3%). Hence, the negative predictive value of the panfungal PCR assay presented is very high, and our current data indicate that molecular screening of patients during febrile neutropenic episodes by the assay can help prevent unnecessary toxicity resulting from empirical antifungal treatment in individuals who may not be at risk of imminent fungal disease. The specificity of the assay in the test cohort of patients was in the range of 76% (95%CI:62-87%), and the observations indicate that rapid species identification may be required to assess the positive predictive value for impending fungus-related disease. The availabel data provide a basis for appropriately designed clinical studies addressing the full diagnostic potential of fungal screening by highly sensitive, broad- spectrum molecular assays in severely immunocompromised patients. Disclosures: No relevant conflicts of interest to declare.

Diagnosis of invasive fungal infections using real-time PCR assay in paediatric acute leukaemia induction

Mycoses ◽  
2012 ◽  
Vol 55 (4) ◽  
pp. 372-379 ◽  
Author(s):  
Sushil Mandhaniya ◽  
Sobuhi Iqbal ◽  
Surender Kumar Sharawat ◽  
Immaculata Xess ◽  
Sameer Bakhshi
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Acute Leukaemia ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Pcr Assay

scholarly journals New Panfungal Real-Time PCR Assay for Diagnosis of Invasive Fungal Infections

2016 ◽  
Vol 54 (12) ◽  
pp. 2910-2918 ◽  
Author(s):  
Clara Valero ◽  
Laura de la Cruz-Villar ◽  
Óscar Zaragoza ◽  
María José Buitrago
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Fungal Infections ◽  
Melting Curve ◽  
Fungal Species ◽  
Invasive Fungal Infections ◽  
Clinical Samples ◽  
Melting Curve Analysis ◽  
Pcr Assay ◽  
Suitable Alternative

The diagnosis of invasive fungal infections (IFIs) is usually based on the isolation of the fungus in culture and histopathological techniques. However, these methods have many limitations often delaying the definitive diagnosis. In recent years, molecular diagnostics methods have emerged as a suitable alternative for IFI diagnosis. When there is not a clear suspicion of the fungus involved in the IFI, panfungal real-time PCR assays have been used, allowing amplification of any fungal DNA. However, this approach requires subsequent amplicon sequencing to identify the fungal species involved, increasing response time. In this work, a new panfungal real-time PCR assay using the combination of an intercalating dye and sequence-specific probes was developed. After DNA amplification, a melting curve analysis was also performed. The technique was standardized by using 11 different fungal species and validated in 60 clinical samples from patients with proven and probable IFI. A melting curve database was constructed by collecting those melting curves obtained from fungal species included in the standardization assay. Results showed high reproducibility (coefficient of variation [CV] < 5%; r > 0.95) and specificity (100%). The overall sensitivity of the technique was 83.3%, with the group of fungi involved in the infection detected in 77.8% of the positive samples with IFIs covered by molecular beacon probes. Moreover, sequencing was avoided in 67.8% of these “probe-positive” results, enabling report of a positive result in 24 h. This technique is fast, sensitive, and specific and promises to be useful for improving early diagnosis of IFIs.

scholarly journals Clinical evaluation of an in-house panfungal real-time PCR assay for the detection of fungal pathogens

Infection ◽  
2020 ◽  
Vol 48 (3) ◽  
pp. 345-355 ◽  
Author(s):  
Iris Camp ◽  
Gabriele Manhart ◽  
Claudia Schabereiter-Gurtner ◽  
Kathrin Spettel ◽  
Brigitte Selitsch ◽  
...  
Keyword(s):  
Clinical Evaluation ◽  
Fungal Pathogens ◽  
Fungal Infections ◽  
Wide Spectrum ◽  
Invasive Fungal Infections ◽  
Diagnostic Tools ◽  
Clinical Samples ◽  
Pcr Assay ◽  
Blood Samples ◽  
Panfungal Pcr

Abstract Purpose Due to an increasing incidence of invasive fungal infections, the availability of reliable diagnostic tools for the fast detection of a wide spectrum of fungal pathogens is of vital importance. In this study, we aimed to conduct an extensive clinical evaluation of a recently published in-house panfungal PCR assay on samples from suspected invasive fungal infections. Methods Overall 265 clinical samples from 232 patients with suspected invasive fungal disease (96 deep airway samples, 60 sterile fluids, 50 tissue biopsies, and 59 blood samples) were included. All samples underwent standard culture-based diagnostics and were additionally analyzed with our panfungal PCR assay. Results Overall, 55.1% of agreement between culture and the panfungal PCR was observed; in 17% of all samples partial concordance was noted, while results between culture and our PCR assay were not in agreement in 27.9%. Our panfungal assay performed better in samples from normally sterile sites, while samples from the deep airways yielded the highest rate of discordant (39.6%) results. In two tissue and three blood samples an invasive pathogen was only detected by PCR while cultures remained negative. Conclusion In combination with routine methods, our panfungal PCR assay is a valuable diagnostic tool. Patients at risk for invasive fungal infections might profit from the reduced time to pathogen identification.

scholarly journals 113 Invasive fungal infections among pediatric patients with hematologic malignancies at KFSH&RC/KFCCC&R

2006 ◽  
Vol 10 ◽  
pp. S63
Author(s):  
I. Bin-Hussain ◽  
F. Al Kordy ◽  
R. Serhan ◽  
A. Al Ahmari ◽  
A. Belgaumi ◽  
...  
Keyword(s):  
Pediatric Patients ◽  
Fungal Infections ◽  
Hematologic Malignancies ◽  
Invasive Fungal Infections

Prevention and monitoring of invasive fungal infections in pediatric patients with cancer and hematologic disorders

2006 ◽  
Vol 48 (1) ◽  
pp. 28-34 ◽  
Author(s):  
Liisa Hovi ◽  
Harri Saxen ◽  
Ulla M. Saarinen-Pihkala ◽  
Kim Vettenranta ◽  
Taru Meri ◽  
...  
Keyword(s):  
Pediatric Patients ◽  
Fungal Infections ◽  
Invasive Fungal Infections ◽  
Hematologic Disorders ◽  
Patients With Cancer

scholarly journals Panfungal PCR Assay for Detection of Fungal Infection in Human Blood Specimens

1998 ◽  
Vol 36 (5) ◽  
pp. 1169-1175 ◽  
Author(s):  
Jo-Anne Van Burik ◽  
David Myerson ◽  
Randall W. Schreckhise ◽  
Raleigh A. Bowden
Keyword(s):  
Fungal Infection ◽  
Whole Blood ◽  
Limit Of Detection ◽  
Fungal Infections ◽  
Small Subunit ◽  
Rrna Gene ◽  
Small Subunit Rrna ◽  
Pcr Assay ◽  
Blood Specimens ◽  
Panfungal Pcr

A novel panfungal PCR assay which detects the small-subunit rRNA gene sequence of the two major fungal organism groups was used to test whole-blood specimens obtained from a series of blood or bone marrow transplant recipients. The 580-bp PCR product was identified after amplification by panfungal primers and hybridization to a 245-bp digoxigenin-labeled probe. The lower limit of detection of the assay was approximately four organisms per milliliter of blood. Multiple whole-blood specimens from five patients without fungal infection or colonization had negative PCR results. Specimens from 11 infected patients had positive PCR results. Blood from three patients with pulmonary aspergillosis had positive PCR results: one patient’s blood specimen obtained in the week prior to the diagnosis of infection by a positive bronchoalveolar lavage fluid culture result was positive by PCR, and blood specimens obtained from two patients 1 to 2 days after lung biopsy and which were sterile by culture were positive by PCR. The blood of four patients with candidemia, three patients with mixed fungal infections, and one patient with fusariosis also had positive PCR signals. The panfungal PCR assay can detect multiple fungal genera and may be used as an adjunct to conventional methods for the detection of fungal infection or for describing the natural history of fungal infection. Further studies are needed to define the sensitivity and specificity of this assay for the diagnosis of fungal infection prior to the existence of other clinical or laboratory indications of invasive fungal infection.

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